双层粒子转染细胞拯救感染性轮状病毒

Rescue of infectious rotavirus from double layer particle

目的从失去外壳结构蛋白无感染性的双层粒子拯救获得具有感染性的轮状病毒粒子,证实轮状病毒双层粒子在细胞内具有感染性。方法将从羊轮状病毒LLR感染的MA-104中制备和纯化轮状病毒双层粒子转染至CV-1细胞,取转染CV-1细胞的培养液用轮状病毒免疫胶体金试剂盒检测证实转染后产生了轮状病毒,将转染细胞培养液接种MA-104细胞鉴定培养液中转染后产生的轮状病毒的感染性。结果轮状病毒双层粒子转染CV-1细胞后细胞产生细胞病变效应,转染细胞的培养液轮状病毒免疫胶体金试剂盒检测结果阳性,转染细胞的培养液接种MA104细胞后也产生典型的细胞病变效应,试剂盒检测也呈阳性。结论从无感染性的纯化羊轮状病毒LLR双层粒子成功地拯救出具有感染性的轮状病毒粒子,证明失去外壳蛋白的轮状病毒双层粒子在细胞内具有感染性。

Objective To recover infectious rotavirus from non-infectious double layer rotavirus particle and to identify the infectivity of double layer rotavirus particle within cell. Methods Double layer particles of rotavirus were prepared and purified from Rotavirus LLR infected MA-104 cells, and were transfected to CV-1 cells. The supernatant of transfected cells was tested by Rotavirus Group A Diagnostic Kit and was inoculated to cultured MA-104 cells to show the infectivity of rotavinls in transfected to CV-1 cells supernatant. Results The transfected CV-1 cells exhibited typical cytopathic effect （CPE） ,and the supernatant of transfected cells was rotavirus positive when tested by Rotavirus Group A Diagnostic Kit. After MA-104 cells were infected by the supematant of transfected CV-1 cells, the MA-104 ceils also produced typical CPE, and the supematant of infected ceils was rotavirus positive when tested by Rotavirus Group A Diagnostic Kit. Conclusion The infectious rotaviruses were successfully rescued from the noninfectious double layer rotavirus particles, and it has been identified that double layer rotavirus particles have the infectivity within cell.